Fig. 7

Kv1.3 channels are expressed by pro-inflammatory DAM and regulate pro-inflammatory DAM genes. a Confirmation of specificity of flow cytometric assay of functional Kv1.3 channels: HEK293 cells were transiently transfected with 1 μg of pRC/CMV (empty vector) or pRC/CMV-mKv1.3 (mouse Kv1.3) after which ShK-F6CA was added (final concentration 100 nM) × 30 min, followed by flow cytometry (3 independent experiments were performed). Electrophysiological confirmation of Kv1.3 current expression was performed by whole-cell patch clamp (data not shown). b Expression of Magenta module markers Kv1.3 and CD45 in subsets of CD11c⁺ DAM in 5xFAD mice (Homeostatic: CD44⁻CXCR4⁻, Magenta/pro-inflammatory-DAM: CD44⁺CXCR4⁻, Yellow/anti-inflammatory-DAM: CXCR4⁺CD44⁻ and double-positive CD44⁺CXCR4⁺ DAM) in adult 5xFAD mice (n = 5 mice, age 6–8 mo). Grey histogram represents isotype control (for CD45) or negative control (unlabeled cells for ShK-F6CA). c Gating of CNS immune cells based on CD11b and CD45 into CD11b⁺CD45^low (resident microglia) and CD11b⁺CD45^high subpopulations (Top). Kv1.3 surface expression in CD45^high and CD45^low subsets of CD11b⁺ cells, is shown below. d Comparison of Kv1.3 expression in CD45^low and CD45^high subsets of CD11b⁺ CNS immune cells in 3 age groups of WT and 5xFAD mice (n = 3 mice/group/time point. Post-hoc statistical tests: *p < 0.05, **p < 0.01, ***p < 0.005. e Comparison of gene expression in acutely isolated CNS immune cells isolated from 6 mo WT mice treated with saline, ShK-223, LPS or LPS + ShK-223 (4 daily i.p. doses, n = 3/group). Selected module markers: Homeostatic: Tmem119, Pro-inflammatory DAM: Il1b, Ptgs2; Anti-inflammatory DAM: Kcnj2, Nceh1, Timp2